12 results on '"Sarah Line Skovbakke"'
Search Results
2. Structure–Function Characteristics and Signaling Properties of Lipidated Peptidomimetic FPR2 Agonists: Peptoid Stereochemistry and Residues in the Vicinity of the Headgroup Affect Function
- Author
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Claes Dahlgren, André Holdfeldt, Sarah Line Skovbakke, Henrik Franzyk, Huamei Forsman, Christina Nielsen, and Michael Gabl
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Agonist ,Chemistry ,medicine.drug_class ,Stereochemistry ,Peptidomimetic ,General Chemical Engineering ,Structure function ,Inflammation ,Peptoid ,General Chemistry ,Formyl peptide receptor 2 ,lcsh:Chemistry ,chemistry.chemical_compound ,lcsh:QD1-999 ,medicine ,medicine.symptom ,Function (biology) - Abstract
Formyl peptide receptor 2 (FPR2) plays important roles in inflammation. In the present study, 20 analogues of the FPR2-selective lipidated α-peptide/β-peptoid agonist Lau-[(S)-Aoc]-[Lys-βNPhe] 6 -NH 2 were generated, which allowed two novel subclasses of more potent FPR2 agonists to be distinguished. Critical factors influencing FPR2 recognition comprise the presence of β-peptoid phenylalanine-like residues (i.e., βNPhe, βNspe, or βNrpe) in the peptidomimetic tail, configuration of the 2-Aminooctanoic acid (Aoc) in the headgroup, and the length of the N-Terminal fatty acid. Intriguingly, a single βNrpe residue in the vicinity of the N-Terminus (i.e., Lau-[(S)-Aoc]-Lys-βNrpe-[Lys-βNPhe] 5 -NH 2 ) proved to increase the agonist potency, whereas the βNspe-containing analogue was a weak FPR2-selective antagonist. Another subclass displaying potent agonism comprised analogues possessing two α-Amino acids vicinal to the headgroup. The optimized FPR2-Activating lipidated peptidomimetics exhibited biased signaling: PLC-PIP 2 -Ca 2+ signaling was activated, but without recruitment of β-Arrestin or induction of chemotaxis. These FPR2-interacting compounds are considered to be useful tools in future studies of receptor-ligand interactions.
- Published
- 2019
3. Studies on acid stability and solid-phase block synthesis of peptide–peptoid hybrids: ligands for formyl peptide receptors
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Iris Perez-Gassol, Anna Mette Hansen, Henrik Franzyk, Simon Bendt Christensen, and Sarah Line Skovbakke
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0301 basic medicine ,Peptidomimetic ,Fluoroacetates ,Clinical Biochemistry ,Peptide ,Ligands ,Biochemistry ,Oligomer ,Peptoids ,03 medical and health sciences ,Residue (chemistry) ,chemistry.chemical_compound ,Solid-phase synthesis ,Side chain ,Immunologic Factors ,Peptide bond ,Chromatography, High Pressure Liquid ,Solid-Phase Synthesis Techniques ,chemistry.chemical_classification ,030102 biochemistry & molecular biology ,Chemistry ,Organic Chemistry ,Peptoid ,Receptors, Formyl Peptide ,Combinatorial chemistry ,030104 developmental biology ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Peptidomimetics - Abstract
α-Peptoids as well as peptide/α-peptoid hybrids and peptide/β-peptoid hybrids constitute major classes of proteolytically stable peptidomimetics that have been extensively investigated as mimetics of biologically active peptides. Representatives of lipidated peptide/β-peptoid hybrids have been identified as promising immunomodulatory lead compounds, and hence access to these via protocols suitable for gram-scale synthesis is warranted to enable animal in vivo studies. Recent observations indicated that several byproducts appear in crude mixtures of relatively short benzyl-based peptide/β-peptoid oligomers, and that these were most predominant when the β-peptoid units displayed an α-chiral benzyl side chain. This prompted an investigation of their stability under acidic conditions. Simultaneous deprotection and cleavage of peptidomimetics containing either α-chiral α- or β-peptoid residues required treatment with strong acid only for a short time to minimize the formation of partially debenzylated byproducts. The initial work on peptide/β-peptoid oligomers with an alternating design established that it was beneficial to form the amide bond between the carboxyl group of the α-amino acid and the congested amino functionality of the β-peptoid residue in solution. To further simplify oligomer assembly on solid phase, we now present a protocol for purification-free solid-phase synthesis of tetrameric building blocks. Next, syntheses of peptidomimetic ligands via manual solid-phase methodologies involving tetrameric building blocks were found to give more readily purified products as compared to those obtained with dimeric building blocks. Moreover, the tetrameric building blocks could be utilized in automated synthesis with microwave-assisted heating, albeit the purity of the crude products was not increased.
- Published
- 2018
4. The Role of Formyl Peptide Receptors for Immunomodulatory Activities of Antimicrobial Peptides and Peptidomimetics
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Huamei Forsman, Henrik Franzyk, Johan Bylund, Sarah Line Skovbakke, and André Holdfeldt
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0301 basic medicine ,Pharmacology ,Innate immune system ,Effector ,Peptidomimetic ,Chemistry ,Phagocytosis ,Antimicrobial peptides ,Inflammation ,Chemotaxis ,Receptors, Formyl Peptide ,Immunity, Innate ,Immunomodulation ,03 medical and health sciences ,030104 developmental biology ,Biochemistry ,Drug Discovery ,medicine ,Animals ,Humans ,Immunologic Factors ,Peptidomimetics ,medicine.symptom ,Receptor ,Antimicrobial Cationic Peptides - Abstract
In recent years, the therapeutic potential of antimicrobial peptides (AMPs) as immunomodulators has become generally accepted. Nevertheless, only very few AMP-based compounds have progressed into clinical trials. This paradox may be explained by the fact, that some of the intrinsic properties of natural peptides, such as proteolytic and oxidative instability, render them inconvenient as therapeutics. Therefore, substantial research efforts have been dedicated to mimic the physico-chemical properties as well as biological activities of AMPs by designing and identifying more stable peptidomimetics displaying analogous immunomodulatory activity profiles. Neutrophils play key roles in host defense as major effector cells in clearance of pathogens by phagocytosis and by regulating other processes of innate immunity as well as by promoting resolution of inflammation. Several aspects of these effects are correlated to their expression of formyl peptide receptors (FPRs) that have been shown to be targets of both natural and synthetic antimicrobial peptides. In the present review recent findings highlighting the role of FPRs in mediating immunomodulatory activities of natural and synthetic AMPs as well as of stabilized peptidomimetics are discussed, and prospects for future development of immunomodulatory therapeutics are presented.
- Published
- 2018
5. The peptidomimetic Lau-(Lys-βNSpe)6-NH2 antagonizes formyl peptide receptor 2 expressed in mouse neutrophils
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Sara K. Lindén, Claes Dahlgren, Huamei Forsman, Ji Ming Wang, Malene Winther, Henrik Franzyk, Michael Gabl, Sarah Line Skovbakke, and André Holdfeldt
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0301 basic medicine ,Pharmacology ,Agonist ,chemistry.chemical_classification ,Formyl peptide receptor ,Peptidomimetic ,medicine.drug_class ,Antagonist ,Peptide ,Biology ,Biochemistry ,Formyl peptide receptor 2 ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,chemistry ,medicine ,Receptor ,030215 immunology ,G protein-coupled receptor - Abstract
The formyl peptide receptor (FPR) gene family has a complex evolutionary history and comprises eight murine members but only three human representatives. To enable translation of results obtained in mouse models of human diseases, more comprehensive knowledge of the pharmacological similarities/differences between the human and murine FPR family members is required. Compared to FPR1 and FPR2 expressed by human neutrophils, very little is known about agonist/antagonist recognition patterns for their murine orthologues, but now we have identified two potent and selective formylated peptide agonists (fMIFL and PSMα2) for Fpr1 and Fpr2, respectively. These peptides were used to determine the inhibition profile of a set of antagonists with known specificities for the two FPRs in relation to the corresponding murine receptors. Some of the most potent and selective antagonists for the human receptors proved to be devoid of effect on their murine orthologues as determined by their inability to inhibit superoxide release from murine neutrophils upon stimulation with receptor-specific agonists. The Boc-FLFLF peptide was found to be a selective antagonist for Fpr1, whereas the lipidated peptidomimetic Lau-(Lys-βNSpe)6-NH2 and the hexapeptide WRW4 were identified as Fpr2-selective antagonists.
- Published
- 2016
6. The Lipidated Peptidomimetic Lau-((S)-Aoc)-(Lys-βNphe)6-NH2 Is a Novel Formyl Peptide Receptor 2 Agonist That Activates Both Human and Mouse Neutrophil NADPH Oxidase
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Christina Nielsen, Iris Perez-Gassol, Anna Karlsson, Michael Gabl, Camilla Josephine Larsen, Huamei Forsman, Claes Dahlgren, Malene Winther, Ji Ming Wang, Sarah Line Skovbakke, André Holdfeldt, and Henrik Franzyk
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Male ,0301 basic medicine ,Agonist ,Neutrophils ,Peptidomimetic ,medicine.drug_class ,Peptide ,Biochemistry ,Formyl peptide receptor 2 ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,medicine ,Animals ,Humans ,Receptors, Lipoxin ,Receptor ,Molecular Biology ,Mice, Knockout ,chemistry.chemical_classification ,NADPH oxidase ,biology ,Lauric Acids ,NADPH Oxidases ,Peptoid ,Cell Biology ,Receptors, Formyl Peptide ,Enzyme Activation ,030104 developmental biology ,chemistry ,biology.protein ,Female ,Peptidomimetics ,Signal transduction ,Signal Transduction ,030215 immunology - Abstract
Neutrophils expressing formyl peptide receptor 2 (FPR2) play key roles in host defense, immune regulation, and resolution of inflammation. Consequently, the search for FPR2-specific modulators has attracted much attention due to its therapeutic potential. Earlier described agonists for this receptor display potent activity for the human receptor (FPR2) but low activity for the mouse receptor orthologue (Fpr2), rendering them inapplicable in murine models of human disease. Here we describe a novel FPR2 agonist, the proteolytically stable α-peptide/β-peptoid hybrid Lau-((S)-Aoc)-(Lys-βNphe)6-NH2 (F2M2), showing comparable potency in activating human and mouse neutrophils by inducing a rise in intracellular Ca(2+) concentration and assembly of the superoxide-generating NADPH oxidase. This FPR2/Fpr2 agonist contains a headgroup consisting of a 2-aminooctanoic acid (Aoc) residue acylated with lauric acid (C12 fatty acid), which is linked to a peptide/peptoid repeat ((Lys-βNphe)6-NH2). Both the fatty acid moiety and the (S)-Aoc residue were required for FPR2/Fpr2 activation. This type of proteolytically stable FPR2-specific peptidomimetics may serve as valuable tools for future analysis of FPR2 signaling as well as for development of prophylactic immunomodulatory therapy. This novel class of cross-species FPR2/Fpr2 agonists should enable translation of results obtained with mouse neutrophils (and disease models) into enhanced understanding of human inflammatory and immune diseases.
- Published
- 2016
7. The proteolytically stable peptidomimetic Pam-(Lys-βNSpe)6-NH2 selectively inhibits human neutrophil activation via formyl peptide receptor 2
- Author
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Henrik Franzyk, Huamei Forsman, Claes Dahlgren, Peter M. H. Heegaard, Sarah Line Skovbakke, and Camilla Josephine Larsen
- Subjects
Agonist ,Neutrophils ,Peptidomimetic ,medicine.drug_class ,Proteolysis ,G-protein-coupled receptors ,HL-60 Cells ,Peptide ,Biochemistry ,Neutrophil Activation ,Formyl peptide receptor 2 ,chemistry.chemical_compound ,In vivo ,medicine ,Humans ,Receptors, Lipoxin ,G protein-coupled receptor ,Pharmacology ,chemistry.chemical_classification ,Dose-Response Relationship, Drug ,medicine.diagnostic_test ,Chemistry ,Host defense peptides ,Receptors, Formyl Peptide ,PyBOP ,Peptidomimetics ,Anti-inflammatory ,Formyl peptide receptors - Abstract
Immunomodulatory host defense peptides (HDPs) are considered to be lead compounds for novel anti-sepsis and anti-inflammatory agents. However, development of drugs based on HDPs has been hampered by problems with toxicity and low bioavailability due to in vivo proteolysis. Here, a subclass of proteolytically stable HDP mimics consisting of lipidated α-peptide/β-peptoid oligomers was investigated for their effect on neutrophil function. The most promising compound, Pam-(Lys-βNSpe)6-NH2, was shown to inhibit formyl peptide receptor 2 (FPR2) agonist-induced neutrophil granule mobilization and release of reactive oxygen species. The potency of Pam-(Lys-βNSpe)6-NH2 was comparable to that of PBP10, the most potent FPR2-selective inhibitor known. The immunomodulatory effects of structural analogues of Pam-(Lys-βNSpe)6-NH2 emphasized the importance of both the lipid and peptidomimetic parts. By using imaging flow cytometry in primary neutrophils and FPR-transfected cell lines we found that a fluorescently labelled analogue of Pam-(Lys-βNSpe)6-NH2 interacted selectively with FPR2. Furthermore the interaction between Pam-(Lys-βNSpe)6-NH2 and FPR2 was found to prevent binding of the FPR2-specific activating peptide agonist Cy5-WKYMWM, while the binding of a FPR1-selective agonist was not inhibited. To our knowledge, Pam-(Lys-βNSpe)6-NH2 is the first HDP mimic found to inhibit activation of human neutrophils via direct interaction with FPR2. Hence, we consider Pam-(Lys-βNSpe)6-NH2 to be a convenient tool in the further dissection of the role of FPR2 in inflammation and homeostasis as well as for investigation of the importance of neutrophil stimulation in anti-infective therapy involving HDPs.
- Published
- 2015
8. Combining Elements from Two Antagonists of Formyl Peptide Receptor 2 Generates More Potent Peptidomimetic Antagonists
- Author
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Henrik Franzyk, Iris Perez-Gassol, Huamei Forsman, Sarah Line Skovbakke, Christina Nielsen, Claes Dahlgren, Anna Mette Hansen, and André Holdfeldt
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0301 basic medicine ,Peptidomimetic ,Stereochemistry ,Neutrophils ,Enzyme Activators ,Peptide ,Subclass ,Formyl peptide receptor 2 ,03 medical and health sciences ,Structure-Activity Relationship ,0302 clinical medicine ,Superoxides ,Drug Discovery ,Structure–activity relationship ,Humans ,Receptors, Lipoxin ,Receptor ,Gelsolin ,chemistry.chemical_classification ,Molecular Structure ,Antagonist ,NADPH Oxidases ,Combinatorial chemistry ,Receptors, Formyl Peptide ,Peptide Fragments ,N-Formylmethionine Leucyl-Phenylalanine ,030104 developmental biology ,chemistry ,Molecular Medicine ,Peptidomimetics ,Peptides ,Oligopeptides ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
Structural optimization of a peptidomimetic antagonist of formyl peptide receptor 2 (FPR2) was explored by an approach involving combination of elements from the two most potent FPR2 antagonists described: a Rhodamine B-conjugated 10-residue gelsonin-derived peptide (i.e., PBP10, RhB-QRLFQVKGRR-OH) and the palmitoylated α-peptide/β-peptoid hybrid Pam-(Lys-βNspe)6-NH2. This generated an array of hybrid compounds from which a new subclass of receptor-selective antagonists was identified. The most potent representatives displayed activity in the low nanomolar range. The resulting stable and potent FPR2-selective antagonists (i.e., RhB-(Lys-βNphe)n-NH2; n = 4–6) are expected to become valuable tools in further elucidation of the physiological role of FPR2 in health and disease.
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- 2017
9. Anti-inflammatory Properties of Antimicrobial Peptides and Peptidomimetics: LPS and LTA Neutralization
- Author
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Sarah Line Skovbakke and Henrik Franzyk
- Subjects
0301 basic medicine ,Lipopolysaccharide ,Chemistry ,Peptidomimetic ,medicine.drug_class ,medicine.medical_treatment ,Antimicrobial peptides ,Pharmacology ,Neutralization ,Anti-inflammatory ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,0302 clinical medicine ,Cytokine ,In vivo ,030220 oncology & carcinogenesis ,medicine ,Lipoteichoic acid - Abstract
Lipopolysaccharide (LPS) and lipoteichoic acid (LTA) neutralization constitute potential non-antibiotic treatment strategies for sepsis - a systemic infection-induced inflammatory response. Studies on LPS- and LTA-neutralizing compounds are abundant in literature, and a number of peptides and peptidomimetics appear to display promising activity. However, in this ongoing search for potential antisepsis drug leads, it will be preferable that the assays used by different research groups lead to readily comparable data for the most efficient compounds. Here, we propose and describe standardized methods to be used for testing of novel compounds for their LPS- and LTA-neutralizing capacity with a focus on functional suppression of pro-inflammatory responses in cell-based systems. To best mimic the human in vivo conditions, we suggest the use of freshly isolated human leukocytes combined with an appropriate method for the chosen cytokine (e.g., IL-6 or TNF-α). The described protocols comprise isolation, stimulation, and viability test of the human leukocytes.
- Published
- 2016
10. Structural changes of the ligand and of the receptor alters the receptor preference for neutrophil activating peptides starting with a formylmethionyl group
- Author
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Huamei Forsman, François Boulay, Michael Gabl, Malene Winther, Marie-Josèphe Rabiet, Claes Dahlgren, Sarah Line Skovbakke, Department of Rheumatology & Inflammation Research, University of Gothenburg (GU), Biology of Cancer and Infection, and Université Joseph Fourier - Grenoble 1 (UJF)
- Subjects
Formyl peptide ,Peptide ,MESH: Receptors, Formyl Peptide ,MESH: Receptors, Lipoxin ,Biology ,Ligands ,Neutrophil Activation ,Formyl peptide receptor 1 ,Structure-Activity Relationship ,03 medical and health sciences ,0302 clinical medicine ,MESH: Structure-Activity Relationship ,MESH: Ligands ,Humans ,5-HT5A receptor ,Receptors, Lipoxin ,Receptor ,Molecular Biology ,Glucagon-like peptide 1 receptor ,030304 developmental biology ,G protein-coupled receptor ,chemistry.chemical_classification ,Inflammation ,0303 health sciences ,MESH: Humans ,Tetrapeptide ,MESH: N-Formylmethionine Leucyl-Phenylalanine ,MESH: Neutrophil Activation ,[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,Neutrophil ,Cell Biology ,Receptors, Formyl Peptide ,N-Formylmethionine Leucyl-Phenylalanine ,chemistry ,Biochemistry ,G-protein coupled receptor ,Relaxin/insulin-like family peptide receptor 2 ,030215 immunology - Abstract
International audience; Pathogenic Staphylococcus aureus strains produce N-formylmethionyl containing peptides, of which the tetrapeptide fMIFL is a potent activator of the neutrophil formyl peptide receptor 1 (FPR1) and the PSMα2 peptide is a potent activator of the closely related FPR2. Variants derived from these two peptide activators were used to disclose the structural determinants for receptor interaction. Removal of five amino acids from the C-terminus of PSMα2 gave rise to a peptide that had lost the receptor-independent neutrophil permeabilizing effect, whereas neutrophil activation capacity as well as its preference for FPR2 was retained. Shorter peptides, PSMα21-10 and PSMα21-5, activate neutrophils, but the receptor preference for these peptides was switched to FPR1. The fMIFL-PSM5-16 peptide, in which the N-terminus of PSMα21-16 was replaced by the sequence fMIFL, was a dual agonist for FPR1/FPR2, whereas fMIFL-PSM5-10 preferred FPR1 to FPR2. Further, an Ile residue was identified as a key determinant for interaction with FPR2. A chimeric receptor in which the cytoplasmic tail of FPR1 was replaced by the corresponding part of FPR2 lost the ability to recognize FPR1 agonists, but gained function in relation to FPR2 agonists. Taken together, our data demonstrate that the C-terminus of the PSMα2 peptide plays a critical role for its cytotoxicity, but is not essential for the receptor-mediated pro-inflammatory activity. More importantly, we show that the amino acids present in the C-terminus, which are not supposed to occupy the agonist-binding pocket in the FPRs, are of importance for the choice of receptor.
- Published
- 2015
11. Lipidated α-peptide/β-peptoid hybrids with potent anti-inflammatory activity
- Author
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Henrik Franzyk, Sarah Line Skovbakke, Camilla Josephine Larsen, Peter M. H. Heegaard, and Lise Moesby
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Lipopolysaccharides ,medicine.drug_class ,Polymyxin ,Anti-Inflammatory Agents ,Peptide ,Flow cytometry ,chemistry.chemical_compound ,Peptoids ,Structure-Activity Relationship ,Drug Discovery ,medicine ,Immunologic Factors ,Cytotoxicity ,chemistry.chemical_classification ,medicine.diagnostic_test ,Interleukin-6 ,Tumor Necrosis Factor-alpha ,Peptoid ,chemistry ,Biochemistry ,Limulus amebocyte lysate ,Molecular Medicine ,Cytokine secretion ,Peptidomimetics ,Polymyxin B ,medicine.drug - Abstract
In this study, we investigated, optimized, and characterized a novel subclass of host defense peptide (HDP) mimics based on α-peptide/β-peptoid hybrid oligomers with an alternating cationic/hydrophobic design with respect to their ability to modulate the pro-inflammatory response by human primary leukocytes upon exposure to bacterial components. Structure-activity studies revealed that certain lipidated α-peptide/β-peptoid hybrid oligomers possess anti-inflammatory activities in the submicromolar range with low cytotoxicity, and that the anti-inflammatory activity of the HDP mimics is dependent on the length and position of the lipid element(s). The resulting lead compound, Pam-(Lys-βNSpe)6-NH2, blocks LPS-induced cytokine secretion with a potency comparable to that of polymyxin B. The mode of action of this HDP mimic appears not to involve direct LPS interaction since it, in contrast to polymyxin B, displayed only minor activity in the Limulus amebocyte lysate assay. Flow cytometry data showed specific interaction of a fluorophore-labeled lipidated α-peptide/β-peptoid hybrid with monocytes and granulocytes indicating a cellular target expressed by these leukocyte subsets.
- Published
- 2014
12. 2-deoxy D-glucose prevents cell surface expression of NKG2D ligands through inhibition of N-linked glycosylation
- Author
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Helle Jensen, Søren Skov, Karen Hansen, G. Persson, Lars Andresen, Sarah Line Skovbakke, and Michael Hagemann-Jensen
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Glycosylation ,medicine.drug_class ,T-Lymphocytes ,Immunology ,Biology ,Deoxyglucose ,Ligands ,chemistry.chemical_compound ,N-linked glycosylation ,Cell Line, Tumor ,medicine ,Immunology and Allergy ,Humans ,RNA, Small Interfering ,Tunicamycin ,HEK 293 cells ,Histone deacetylase inhibitor ,Histocompatibility Antigens Class I ,Membrane Proteins ,Fusion protein ,Molecular biology ,Histone Deacetylase Inhibitors ,Killer Cells, Natural ,stomatognathic diseases ,HEK293 Cells ,chemistry ,Cell culture ,NK Cell Lectin-Like Receptor Subfamily K ,RNA Interference ,2-Deoxy-D-glucose ,Glycolysis ,Mannose - Abstract
NKG2D ligand surface expression is important for immune recognition of stressed and neotransformed cells. In this study, we show that surface expression of MICA/B and other NKG2D ligands is dependent on N-linked glycosylation. The inhibitor of glycolysis and N-linked glycosylation, 2-deoxy-d-glucose (2DG), potently inhibited surface expression of MICA/B after histone deacetylase inhibitor treatment; the inhibition occurred posttranscriptionally without affecting MICA promoter activity. Transient overexpression of MICA surface expression was also inhibited by 2DG. 2DG blocks N-linked glycosylation of MICA/B by a reversible mechanism that can be alleviated by addition of d-mannose; this does not, however, affect the inhibition of glycolysis. Addition of d-mannose restored MICA/B surface expression after 2DG treatment. In addition, specific pharmacological or small interfering RNA-mediated targeting of glycolytic enzymes did not affect MICA/B surface expression, strongly suggesting that N-linked glycosylation, and not glycolysis, is essential for MICA/B surface expression. Corroborating this, tunicamycin, a selective inhibitor of N-linked glycosylation, abolished MICA/B surface expression without compromising activation of MICA promoter activity. NK cell-mediated killing assay and staining with a recombinant NKG2D–Fc fusion protein showed that all functional NKG2D ligands induced by histone deacetylase inhibitor treatment were abolished by 2DG treatment and fully reconstituted by further addition of d-mannose. Our data suggest that posttranslational N-linked glycosylation is strictly required for NKG2D ligand surface expression. Cancer and infection often result in aberrant glycosylation, which could likely be involved in modulation of NKG2D ligand expression. Our data further imply that chemotherapeutic use of 2DG may restrict NKG2D ligand surface expression and inhibit secretion of immunoinhibitory soluble NKG2D ligands.
- Published
- 2012
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